| 补体C5a受体拮抗剂体外筛选平台的建立 |
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| 引用本文： | 王会会,陈国江,杨跃梅,魏化伟,王立燕,彭晖,李新颖,王一,黎燕,石艳春. 补体C5a受体拮抗剂体外筛选平台的建立[J]. 国外医学(药学分册), 2011, 0(1) |
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| 作者姓名： | 王会会 陈国江 杨跃梅 魏化伟 王立燕 彭晖 李新颖 王一 黎燕 石艳春 |
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| 作者单位： | 内蒙古医学院分子生物学中心;军事医学科学院基础医学研究所分子免疫学研究室; |
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| 基金项目： | 国家自然科学基金资助项目(30801029); 国家重点基础研究发展计划(973计划)项目(2007CB512406); 内蒙古科技计划项目(20080502); 内蒙古自然科学基金项目(2009MS1102) |
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| 摘 要： | 目的建立稳定的补体C5a受体(C5aR)拮抗剂筛选平台。方法采集人外周静脉抗凝血,与不同浓度C5a、脂多糖及PMX-53孵育10～30 min。通过流式细胞术检测中性粒细胞表面CD11b的表达,溶菌酶检测试剂盒观察中性粒细胞分泌溶菌酶的能力变化,罗丹明-123检测中性粒细胞呼吸爆发的变化,ELISA检测细胞上清液中白细胞介素(IL)-8的表达变化,Western Blotting检测胞外信号调节激酶(ERK)、蛋白激酶B(AKB或AKT)含量及其磷酸化水平的变化,考察补体C5a刺激及使用阳性药物PMX-53后对各生化指标的影响。结果 C5a刺激人外周血可增强中性粒细胞CD11b的表达,促进溶菌酶释放和IL-8的分泌,激发呼吸爆发,上调ERK、AKT的含量和磷酸化水平,阳性药物PMX-53则能显著抑制C5a的上述生物学效应。结论成功建立C5aR拮抗剂人全血体外筛选平台,为C5aR拮抗剂筛选及功能研究奠定了基础。
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| 关 键 词： | C5a CD88 C5a受体拮抗剂 筛选平台 PMX-53 |
Establishment of screening platform in vitro for C5a receptor antagonists |
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| WANG Hui-hui,CHEN Guo-jiang,YANG Yue-mei,WEI Hua-wei,WANG Li-yan,PENG Hui,LI Xin-yin,Wang Li,Li Yan,SHI Yan-chun. Establishment of screening platform in vitro for C5a receptor antagonists[J]. Foreign Medical Sciences(Section of Pharmarcy), 2011, 0(1) |
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| Authors: | WANG Hui-hui CHEN Guo-jiang YANG Yue-mei WEI Hua-wei WANG Li-yan PENG Hui LI Xin-yin Wang Li Li Yan SHI Yan-chun |
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| Affiliation: | WANG Hui-hui1,2,CHEN Guo-jiang2,YANG Yue-mei2,WEI Hua-wei2,WANG Li-yan2,PENG Hui2,LI Xin-yin2,Wang Li2,Li Yan2,SHI Yan-chun1(1.Research Center of Molecular Biology,Inner Mongolia Medical College,Hohhot 010059,China,2.Department of Molecular Immunology,Institute of Basic Medical Sciences,Academy of Military Medical Sciences,Beijing 100850,China) |
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| Abstract: | Objective To establish the stable methodology for screening antagonists of CD88,one of C5a receptors.Methods Whole blood from volunteers was collected and then stimulated by C5a with different concentration for 10-30 min in the presence or absence of PMX-53 and LPS.Flow cytometry was used to detect the expression of CD11b which on the surface of neutrophil.Other functions of nentrophil,including lysozyme release,oxidative burst and interleukin(IL)-8 production,were also examined respectively.Finally,western... |
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| Keywords: | C5a CD88 C5a receptor antagonist screening platform PMX-53 |
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